MC-002 exhibits positive effects against platelets aggregation and endothelial dysfunction through thromboxane A2 inhibition

Introduction: Thromboxane A(2) (TXA(2)) induces platelet aggregation and vasoconstriction, and agents that inhibit TXA(2) production or interaction with receptors may exert potential application in stroke therapy. Aim: To illustrate the platelet aggregation antagonistic and endothelial protective effect of (E) - 3 - (3 - methoxy 4 - ((3, 5, 6 - trimethylpyrazin - 2 - yl) methoxy) phenyl) sodium acrylate (MC-002) through TXA(2) inhibition and underline mechanisms. Materials and methods: Platelets aggregation and thoracic aorta ring contraction of rabbits were induced by U46619. Human umbilical vein endothelial cells (HUVECs) were further applied to explore the protective effect of MC-002 on endothelium when exposed to tumor necrosis factor - alpha (TNF-alpha). MTT method was used to assess cell damage, and ELISA analysis was exerted to estimate nitrogen monoxide (NO), endothelin-1 (ET-1), thromboxane B-2 (TXB2) and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) releasing. Fluorescence spectrophotometry was conducted to determine intracellular calcium concentration ([Ca2+](i)), and western blotting method was applied to evaluate the protein expressions of intracellular adhesion molecule-1 (ICAM-1), P-selectin and nuclear factor-kappa B (NF-kappa B). Results and conclusions: TXA(2) analog U46619 mediated obvious platelet aggregation and vasoconstriction. MC-002 inhibited platelet aggregation through administration in vivo and incubation with platelet in vitro, and relaxed aorta ring in endothelium dependent manner. MC-002 alleviated cell damage, [Ca2+] i overload, ET-1 overexcretion and TXB2 activation, but improved NO availability reduction in HUVECs treated with TNF-alpha. Furthermore, MC-002 downregulated ICAM-1, P-selectin and NF-kappa B overexpression induced by TNF-alpha. In conclusion, MC-002 exerted antiplatelet aggregation effect through TXA(2) inhibition and relieved inflammatory injury of endothelial cells through NF-kappa B signal pathway. (C) 2014 Elsevier Ltd. All rights reserved.